DNA-damaging agents typically react with DNA to form a plethora of adducts within the genome. Historically, it has been difficult to ascertain the genetic effects of individual DNA adducts by studying randomly modified DNAs. Over the past decade techniques that allow the precise evaluation of the biological effects of single DNA adducts have been developed. This paper describes the methodology required to determine the mutagenic effects of a single DNA adduct in a biological system. The synthesis and characterization of singly modified oligonucleotides, the incorporation of modified oligonucleotides into biologically viable viral or plasmid genomes, the replication of the singly modified vectors in vivo to allow the fixation of mutations, and subsequent qualitative and quantitative evaluation of adduct-induced mutations are described.
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)